Frontiers in Stem Cells and Regeneration Advanced Training Course September 25 - October 1, 2016 MBL • Woods Hole, MA

Course Directors: Gerald Schatten, PhD Jennifer Morgan, PhD Pittsburgh Development Center Marine Biological Lab

Organizing Committee: Diane Carlisle, PhD; Gloria Perez, DVM; Dorianne Mebane MBA

Faculty Evangelists: Susan Bryant, PhD; Jose García-Arrarás, PhD; David Gardiner, PhD; Malcolm Maden, PhD

All lectures are in Loeb G70 (south), unless otherwise specified. Lab exercises are held in Loeb 306. Meals are in the Swope dining room, 2nd floor. Breakfast: 7-8:30 AM; Lunch: 11:30 AM-1 PM; Dinner: 5-7 PM. Coffee breaks will be served in the Loeb ground floor corridor adjacent to the lecture hall.

Sunday, September 25

5:30 PM Dinner, Café Swope (first floor)

6:30 PM Lecture: Introduction to Regeneration Ken Muneoka, Texas A&M

7:15 PM Lecture: Challenges in Stem Cells and Regeneration Jerry Schatten, U. Pittsburgh

8:00 PM EEO Information and Logistics: Biosafety, Regulatory Restrictions, MOUs, and Team Assignments Diane Carlisle, U. Pittsburgh

Monday, September 26

8:30 AM MBL Welcome Rae Nishi, MBL Director of Education

8:45 AM Lecture: Stem Cells, Pluripotency and Reprogramming Andras Nagy, Lunenfeld-Tanenbaum Research Institute, Toronto

9:30 AM Lecture: Directed Differentiation and hESC as Developmental Sensors Diane Carlisle, U. Pittsburgh

10:15 AM Coffee Break

10:30 AM Lecture: Patient Specific Pluripotent Stem Cells Charles Easley, Emory

11:15 AM Lab Introduction: IPS Cell Derivation, Culture, and Differentiation Charles Easley

11:30 AM Lab Introduction: Calvin Simerly - mESC derivation

Noon Lunch

1:00 PM Lab 1: Instructor: Chas Easley. TA: Danielle Clarkson-Townsend (Emory). Students will learn how to generate human iPSCs using Cytotune 2.0 (Sendai virus) and perform the initial steps of reprogramming. Students will also learn how to isolate colonies from reprogrammed lines and then learn how to passage established human iPSCs/ESCs.

Lab 2: Instructor: Cal Simerly. TA: Kristin Fowler (Tulane). Mouse embryonic (mESC) derivation: Students will do immunosurgery technique on mouse expanded blastocysts to isolate the ICM in order to derive mESCs.

6:00 PM Dinner

7:00 PM Bioethics Lecture Pasquale Patrizio, Yale

8:00 PM Course Participants’ Introductions

Tuesday, September 27

8:30 AM Lecture: Growth Control of Regenerating Limbs James Monaghan, Northeastern University

9:15 AM Lecture: Molecular Basis of Limb Regeneration Randal Voss, U Kentucky

10:00 AM Coffee Break

10:15 AM Lecture and Lab Intro: Stem cells and Regeneration in Planarians David Forsthoefel, OK Medical Research Foundation

11:15 AM Lab Introduction: David Gardiner and Randall Voss - Salamander regeneration

11:30 AM Special Topics Lunch – with vendor sponsors

1:00 PM Lab 1: Instructors: James Monaghan. TA: Johanna Farkas (Northeastern); Regeneration in Salamanders: limb regeneration

Lab 2: Instructor: David Forsthoefel, U. Illinois Regeneration in Planarians

6:00 PM Dinner

2 7:00 PM Course Participants’ Introductions

Wednesday, September 28

8:30 AM Lecture: Picking your favorite - cell identification and selection strategies in human neural stem cell Jan Pruszak, U. Freiburg

9:15 AM Lecture: Spinal Cord Regeneration in a Non-Mammalian Model Jen Morgan, MBL

10:00 AM Coffee Break

10:15 AM Lecture: Discovering gene regulatory networks that drive successful CNS regeneration in zebrafish Ava Udvadia – Univ. of Wisc. Milwaukee

11:00 AM Lab Introduction: Jan Pruszak - Isolating and Characterizing Neural stem cells using FACS strategy.

11:30 AM Lab Introduction: Jen Morgan - Lamprey model of spinal cord injury and regeneration

Noon Lunch

1:00 PM Lab 1: Instructor: Jan Pruszak. In this laboratory, trainees will learn the techniques for isolating specific PSC-derived subsets of therapeutic and scientific relevance using flow cytometry.

Lab 2: Instructor: Jen Morgan. TA: Kendra Hanslik (MBL). Students will learn to quantify behavioral recovery after spinal cord injury and will examine various aspects of nervous tissue regeneration (e.g. axons, neurons, glial cells).

6:00 PM Dinner

7:00 PM Discussion: Stem Cell and Regeneration Research Funding Ravi Ravindranath, NIH

8:00 PM Career Mentoring Discussion Facilitated by Jennifer Morgan (MBL) and other course faculty (*Please bring a copy of your CV)

Thursday, September 29

8:30 AM Lecture: Regeneration and Reading and Writing the Morphogenetic Code Michael Levin, Tufts University

9:15 AM Lecture: Translating Spermatogonial Stem Cell

3 Transplantation to the Clinic Kyle Orwig, U. Pittsburgh

10:00 AM Coffee Break

10:10 AM Lab Intro: Directed differentiation Diane Carlisle, U. Pittsburgh

10:35 AM Lab Intro: Spermatogonial Stem Cell Transplantation Kyle Orwig, U. Pittsburgh

11:00 AM Orwig Lab Prep: Differential separation of cells and apply antibodies for staining Carlisle Lab Prep: Preparing differentiated cells for IF staining

Noon Lunch

1:00 PM Lab 1: Instructor: Diane Carlisle. TA: Marielle Siebert (U. Pittsburgh). We will use differential immunocytochemistry staining to identify enriched populations in cells after directed differentiation.

Lab 2: Instructor: Kyle Orwig. TAs: Meena Sukhwani (U. Pittsburgh), Sherin David (U. Pittsburgh). Germ stem cell isolation, identification, and transplantation using a magnetic sorting (MACS) strategy.

6:00 PM Dinner

7:00 PM open

8:30 PM T-Shirt Night Celebration

Friday, September 30

8:30 AM Lecture: Mechanisms of epigenetic regulation in stem cells and development Alexander Meissner, Harvard Stem Cell Institute

9:15 AM Lecture: Preimplantation Genetic Diagnosis Mark Hughes, Genesis

10:00 AM Coffee Break

10:15 AM PIONEER LECTURE: Neural Stem Cells Arturo Alvarez-Buylla, UCSF

11:30 AM Lab Introduction for ongoing experiments: Kyle Orwig, Charles Easley, Calvin Simerly

11:45 AM Course Photo 4

Noon Lunch

1:00 PM Lab: Finish all ongoing experiments with all faculty: Instructor: Chas Easley: Analysis of iPSCs using . Check on your hESC culture. Instructor: Cal Simerly. Check on mESC derivation. Instructor: James Monaghan. Image blastemas.

3:00 PM Optional: Tour of Marine Resources Center and MBL

5:00 PM PIONEER LECTURE: Epithelial Stem Cells Elaine Fuchs, Rockefeller

6:30 PM Banquet: Café Swope

8:00 PM Presentation of Pioneer Awards and Student Certificates Jennifer Morgan, Gerald Schatten

Saturday, Oct 1

Check out of rooms before 10:00 AM

Thank you to the following course sponsors who have contributed to the course by loaning equipment and/or donating reagents and supplies:

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